Bacillus anthracis, Yersinia pestis, Francisella tularensis, Brucella sp., Bulkholderia mallei are B. pseudomallei are highly pathogenic bacteria of potential bioterrorism risk. To support the early warning and rapid response capacity to ensure an effective reaction to bioterrorist attacks the international project "Establishment of Quality Assurances for Detection of Highly Pathogenic Bacteria of Potential Bioterrorism Risk" (EQADeBa) has been established. The aim of the project was establishment the network of European laboratories that possess a suitable infrastructure and experts and laboratory workers trained in detection and identification of the highly pathogenic bacteria. This work presents methods used in investigation of samples containing living pathogens and results obtained in Department of Bacteriology NIZP-PZH in the third international test organized within the framework of the EQADeBa. The test evaluated competence of laboratories in detection and identification of pathogens mentioned above. In the work methods used in other laboratories were discussed and the results were compared. Results of the test confirmed competence of NIZP-PZH in detection and identification of highly pathogenic bacteria covered by EQADeBa project.